1.Isolation of human genomic DNA from archival dried blood spots for neonatal disease screening and its application to methylation detection
Qiuping CHEN ; Xianwei CUI ; Lianghui YOU ; Chenbo JI ; Xirong GUO
Chinese Journal of Applied Clinical Pediatrics 2015;30(8):615-618
Objective To establish an effective DNA isolation method for neonatal disease screening,so as to explore its application to the methylation detection.Methods The 20 dried blood spots samples were randomly divided into 2 groups according to the gender:the traditional method group (n =10) and the improved kit method group(n =10).The DNA quality was evaluated based on its concentration,integrity and whether it could be used in polymerase chain reaction (PCR).These DNA samples with or without bisulfite treatment were used as template in the methylation-specific polymerase chain reaction (MSP).The methylation levels of Leptin and tumor necrosis factor-α (TNF-α) gene promoter region were detected.Results DNA concentration of the improved kit method [(5.70 ± 0.81) mg/L] was significantly higher than that of the traditional method [(3.50 ± 0.45) mg/L] (t =2.79,P < 0.05),and biochemical analyzer analysis showed a better DNA integrity.Agarose gel electrophoresis revealed that 18S gene fragment could be successfully amplified by PCR method,suggesting its potential application to PCR study.MSP results showed different DNA methylation levels of Leptin and TNF-α genes promoter regions from various samples.Conclusions The improved kit method can effectively extract DNA from dried blood spots samples,and these DNA can be used in methylation research.The study can provide a new research direction and technical method to reveal the pathogenesis of disease from the perspective of DNA methylation.
2.Development of the animal experiment in cartilage tissue engineering and its clinical application
You CHENG ; Tianyou WANG ; Zeqing LI ; Qiuping WANG
Journal of Medical Postgraduates 2003;0(12):-
At present,the animal experiment in cartilage tissue engineering has entered the stage of maturely immune mammal,and has been applied to clinical practice rudimentarily with good prospects,as well as with many difficulties.Although still mainly in the phase of animal experiments,cartilage tissue engineering provided a new means of cartilage defect repairing,maybe the chief direction of research in this domain.This study reviews the current situation and research development of the animal experiment in cartilage tissue engineering and its clinical applications.
3.Effects of allergic and nonallergic rhinitis on bronchial inflammation
Wei CHEN ; Yong ZHANG ; Zhiyi WANG ; Kunmin WU ; Junfeng JI ; You CHENG ; Qiuping WANG ; Tianyou WANG
Journal of Medical Postgraduates 2015;(2):178-181
Objective To describe the effects of allergic and nonallergic rhinitis on patients with bronchial inflammation by comparing the bronchial inflammation characteristics . Methods All patients enrolled in the study were categorized into two groups according to rhinitis diagnostic criteria , 377 cases in the allergic rhinitis ( AR) group, 262 cases in non-allergic rhinitis ( NAR) group and another 264 patients without rhinitis as control group .Induced sputum differential cytology tests , exhaled nitric oxide concentration measurements and methacholine bronchial provocation tests were performed .The characteristics and differences in lower airway inflam-mation among the three groups were compared . Results The exhaled nitric oxide level and the positive rate of the exhaled nitric ox-ide were different significantly in AR group and NAR group than the control group (P<0.05);Bronchial provocation positive rate was 12.20%in AR group, and 6.10%in the NAR group, the difference was significant (P<0.05), and the levels in the two groups were significantly higher than the control group (P<0.05).AR group and NAR group has a significant different (P<0.05) propor-tions of eosinophil in induced sputum , while the positive rate of eosinophil in induced sputum was not different significantly ( P >0.05). Conclusion Bronchial inflammation may be aggravated by AR and NAR , so all patients with AR and NAR need treatment , and the degree of inflammation was higher in AR patients than the NAR patients .
4.Status of patients with allergic rhinitis combined with bronchial asthma in Nanjing and the correlation of clinical characteristics
Fei XUE ; Zeqing LI ; Qiuping WANG ; Xiaofeng XIN ; Manjie JIANG ; You CHENG ; Junfeng JI
Chinese Journal of Tissue Engineering Research 2007;11(43):8809-8812
BACKGROUND: Both Allergic rhinitis and bronchial asthma are allergic diseases of respiratory tract. They frequently coexist in the same patients.OBJECTIVE: To study the incidence of allergic rhinitis in patients with bronchial asthma in Nanjing, and to analyze the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.DESIGN: The questionnaire survey.SETTING: Department of Otorhinolaryngology-Head and Neck Surgery and Department of Respiratory, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.PARTICIPANTS: A total of 134 patients with bronchial asthma including 73 males and 61 females were selected form Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from February 2001 to April 2005. Their ages ranged from 3 to 72 years. Severity of asthma of all patients was classified based on Global Initiative for Asthma (GINA). The diagnostic criteria of allergic rhinitis were established referred to the guideline of Haikou Meeting (November, 1997). All patients provided the confirmed consent.METHODS: The questionnaire survey of 134 bronchial asthma patients from the city of Nanjing was conducted. Their age, classification, severity, treatment status, and the family history of all 134 patients were investigated in details. Differences of age, age at onset, and course of disease between rhinitis and asthma of all patients were examined by two independent t tests. The correlation between clinical characteristics of allergic rhinitis and bronchial asthma was analyzed by Spearman rank correlation.MAIN OUTCOME MEASURES : Incidence of allergic rhinitis in patients with bronchial asthma and the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.RESULTS:Of all 134 bronchial asthma patients,82(61.2%,82/134)were accompanied with allergic rhinitis.There were 45 males and 37 females.Of 82 patients with allergic rhinitis accompanying with bronchial asthma,severity was classified into grade Ⅰ (mild pause,n=56),grade Ⅱ (mild persistence,n=21),grade Ⅲ (moderate persistence,n=4),and grade Ⅳ (severe persistence,n=1).In addition,allergic rhinitis was classified into periodicity (n=65) and persistence(n=17).According to the classification and seventy system recommended in ARIA,63 patients had mild symptoms and 19 had moderate and severe symptoms.Severity of allergic rhinitis was positively related to that of bronchial asthma(r=0.689,P<0.01).CONCLUSION: The morbidity of allergic rhinitis combining with bronchial asthma is high in Nanjing, China. While, their classification and severity are highly coherent.
5.Hydrophilicity and adsorptivity of a novel scaffold twice embedded by lecithin and poly-L-lysine to human nasal septum chondrocytes
You CHENG ; Qiuping WANG ; Manjie JIANG ; Kunmin WU ; Fei XUE ; Wei CHEN ; Junfeng JI ; Zeqing LI
Chinese Journal of Tissue Engineering Research 2007;11(9):1786-1789
BACKGROUND:Chitosan is a kind of natural biomaterial and is characterized by great biocompatibility, progressive degeneration and absorption and excellent mechanical property; however, whether it may become an ideal cytoskeleton in the engineering of cartilage tissue or not should be researched further.OBJECTIVE: To observe the hydrophilicity and adsorptivity to human nasal septum chondrocytes and the effect of its function of a novel scaffold made by [poly (dl-lactide-co-glycolide)] (PLGA)/chitosan nonwoven cloth embedded with lecithin (LEC) and poly-L-lysine (PLYS).DESIGN: Blank control study.SETTING: Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from October 2005 to June 2006. Chitosan nonwoven cloth was provided by Hainan Xinlong Company. The mainly technical parameters were detailed as the follows: degree of deacetylation ≥ 90% and relative molecular weight 2-5 × 105. PLGA/chitosan nonwoven cloth scaffold was made in Department of Otolaryngology-Hean and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA and High Polymer Institute of Sun Yat-sen University. The mainly technical parameters were detailed as the follows: mole ratio between monome lactide and glycolide 75:25, porosity 82%-86%, pore diamater 100-300 μm, shear strength 48 MPa, depth 1.5 mm and completely degenerated duration 14-18 weeks. Human nasal septum chondrocytes were the septal cartilage of nose which was derived from operated patients with deflection of nasal septum under sterile condition.METHODS: PLGA/chitosan nonwoven cloth was sheared into pieces with the size of 1.5 cm × 1.5 cm, dipped in 0.01 volume fraction of LEC anhydrous alcohol and 1 g/L PLYS for 6 hours, dealt with ultraviolet radiation after dehydration for 1hour, dipped in 0.75 volume fraction of ethanol for 24 hours, washed with Hanks solution, and incubated for 24 hours. After operations mentioned above, two kinds of novel scaffolds containing various components were obtained, and they were simple scaffold and scaffold embedded with LEC and PLYS. Cells derived from the third generation of human nasal septum chondrocytes were used to make suspension. In addition, cell suspension was grown on those two scaffolds to determine the degree of hydrophilicity through observing diffused degree of cell suspension. Whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not were observed under phase contrast microscope so as to determine adsorptivity between cells and scaffolds; meanwhile, growth of cells and production of matrix were also observed.MAIN OUTCOME MEASURES: ① Hydrophilicity of those two kinds of scaffolds (diffused degree of cell suspension of human nasal septum chondrocytes on scaffold) and adsorptivity to cells (whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not); ② growth of cells and production of matrix.RESULTS: ① When simple scaffold was put in cell suspension of human nasal septum chondrocytes, cell suspension showing like balls attached to the surface of scaffold, and then, scattered into space of scaffold gradually. During the period of culture, phase contrast microscope indicated that masses of cells attached to the surface of fiber of scaffold.When the petri dish was shaken, cell groups drifted irregularly. The adherent rate was (21±3.7)%. With the cultured time passing by, matrix was not produced. ② When scaffold embedded with LEC and PLYS was put in cell suspension of human nasal septum chondrocytes, cell suspension scattered into space of scaffold rapidly. The adherent rate was (89±5.6)%, which was higher than that of single scaffold group. This suggested that scaffold showed a strong hydrophilicity.Phase contrast microscope indicated that chondrocytes as the form of monome or community were distributed between scaffold and fiber averagely and attached to the surface of fiber of scaffold. When the petri dish was shaken, cell groups did not drift irregularly, and only a few of cells were scarred at the bottom of petri dish. This suggested that scaffold had a strong adsorptivity to cells. One week after culture, matrix showing like cobweb was produced among fibers of scaffold.With the cultured time passing by, matrix was produced abundantly.CONCLUSION: The novel scaffold of PLGA/chitosan nonwoven cloth embedded with LEC and PLYS is characterized by an excellent hydrophilicity and adsorptivity to human nasal septum chondrocytes; meanwhile, it also can secrete matrix.
6.Fabrication of Allogenic Cartilage in A Tube Lined with Epithelium and A Novel Scaffold
You CHENG ; Jinzhong HUANG ; Qiuping WANG ; Zeqing LI ; Tianyou WANG ; Manjie JIANG ; Mei ZHOU
Journal of Audiology and Speech Pathology 2010;18(1):59-62
Objective To study the feasibility of engineering cartilage tissue in a tube lined with epithelium and implanting allogenic chondrocytes into a novel scaffold consisting of chitosan nonwoven cloth coated with poly (DL-lactide-co-glycolide)(PLGA).The focus of the present study is to explore a new way of repairing laryngeal and tracheal defects.Methods Allogenic chondrocytes were obtained from the auricles of 1-month-old rabbits.After being cultured in vitro for three to four passages,the cells were implanted into the scaffolds to form composite grafts and then transplanted into the rabbits.After 6,12,and 18 weeks,the general,histological characteristics were investigated.Results The cobweb-like matrix was observed approximately 1 week after the chondrocytes had been implanted into the scaffolds.At 6 weeks,the matrix was secreted,and there were immature chondrocytes in the grafts.At 12 weeks,the allogenic cartilage in the tube lined with epithelium had been created.Chondrocytes were almost mature and the lacunae had formed.At 18 weeks,the neocartilage was similar to native cartilage.Conclusion It is feasible to fabricate allogenic cartilage in a tube lined with epithelium by implanting allogenic chondrocytes into a novel scaffold made of chitosan nonwoven cloth coated with PLGA.
7.The modified method for human nasal septum chondrocyte culture and the study of biological characters of the chondrocyte
You CHENG ; Qiuping WANG ; Jinghong LI ; Jiang DU ; He QIN ; Qiao TANG ; Jinzhong HUANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(01):-
OBJECTIVE To established a viable and convenient method for human nasoseptal chondrocyte culture in vitro and study the biological characters of the chondrocyte. METHODS The chondrocytes were obtained by incomplete enzyme digestion of cartilage and were seeded with small piece of cartilage in 25 ml culture flasks coated with polylysine previously. This study determines the biologic synthesis of the chondrocytes by toluidine blue and type Ⅱcollagen immunohistochemistry reaction. The proteoglycan synthesis efficiencies of human nasoseptal chondrocytes cultured in vitro were measured by using 35S-Na2SO4 incorporation. The modified method and the conventional method in proteoglycan synthesis were also compared. RESULTS The results suggested that human nasoseptal chondrocytes cultured by modified method still show the normal structure and morphology, and maintain the function of metabolism and self duplication. With the modified method, we can get a higher cell survival rate and more cells. CONCLUSION The modified method is a viable and convenient method.
8.Survey on oral health status of inpatients with schizophrenia
Qiuping YOU ; Wei ZHENG ; Lirong TANG ; Xiuqin ZHAO ; Wenxiu LI ; Yawen LI
Chinese Journal of Postgraduates of Medicine 2015;38(z1):48-51
Objective To investigate and analyze oral health status of inpatients with schizophrenia.Method We selected 105 patients from Beijing Anding hospital included hospitalization length 1-5 years group (39 cases) and hospitalization length over 5 years (55 cases) were investigated with Self-made questionnaire and assessment guide for psychiatric care (OAG-PC).Results Tooth loss and oral health status of schizophrenia inpatients were both poor.There was significant difference in the number of people with tooth loss (x2 =7.68,P < 0.05) and mean number of tooth loss (F =9.20,P < 0.05) between two groups.The OAG-PC total score (F=11.1,P< 0.05),odour from the mouth (F=4.34,P< 0.05),saliva (F=3.93,P< 0.05),mucous membranes (F=5.64,P< 0.05),teeth or dentures (F=5.34,P< 0.05),calculus on teeth (F=15.34,P < 0.05),looseness of teeth (F =14.14,P < 0.05),and No.of teeth in the lower jaw (F =7.70,P < 0.05) in the schizophrenia inpatients with hospitalization length 1-5 years group was better than those with hospitalization length over 5 years,there were significant difference between two groups.There were no significant difference in lips (F =0.69,P > 0.05),tongue (F =0.65,P > 0.05),gums (F =1.87,P > 0.05),appearance of teeth (F=1.52,P> 0.05),and No.of teeth in the upper jaw (F=3.58,P > 0.05) between two groups.Conclusions Patients with schizophrenia are special vulnerable people.The more course of the disease was longer,the more the oral health status was worse.Tooth loss,calculus,and looseness of teeth are much more serious and will influence chew function seriously.Society,medical institutions,and families should pay more attention to the oral health status of patients with schizophrenia.And taking the preventative measures against poor oral health can improve quality of life of patients with schizophrenia.
9.The latest study and development of cartilage tissue engineering.
You CHENG ; Zeqing LI ; Qiuping WANG
Journal of Biomedical Engineering 2007;24(5):1196-1199
In this paper are reviewed the current situation and research progress of cartilage tissue engineering, including seeding cell, three-dimensional cytoskeleton, cell culture system and evaluation of artificial cartilage behavior, animal model, and clinical trial.
Animals
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Cartilage
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cytology
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physiology
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Cell Culture Techniques
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methods
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Cell Differentiation
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Chondrocytes
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cytology
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physiology
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Humans
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Mesenchymal Stromal Cells
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cytology
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physiology
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Models, Animal
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Tissue Engineering
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methods
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trends
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Tissue Scaffolds
10.Changes in small airway function in patients with non-allergic rhinitis
Junfeng JI ; Yong ZHANG ; Li XU ; Fei XUE ; Minghai WU ; Tianyou WANG ; You CHENG ; Manjie JIANG ; Qiuping WANG
Journal of Medical Postgraduates 2015;(9):940-943
Objective Small airway hypofunction is an early manifestation of asthmatic airway injury and is found in patients with non-asthma allergic rhinitis.However, no report has been seen on the changes of small airway function in patients with non-aller-gic rhinitis ( NAR) .This study was to investigate the possibility of small airway lesion in NAR patients and its relationship with airway responsiveness by observing the changes of small airway function in NAR patients without asthma and/or lower airway symptoms. Methods We recruited 324 subjects for this study, including 262 NAR patients and 62 healthy controls, and assigned them to an air-way hyperresponsiveness (AHR) and a non-airway hyperresponsiveness (nAHR) group.All the subjects underwent medical history collection, nasal examination, allergen skin prick test, blood routine test, serum total immunoglobin E assay, pulmonary function test, and bronchial challenge test. Results Compared with the healthy con-trols, the NAR patients showed remarkably lower predicted percenta-ges of maximal mid-expiratory flow ([85.6 ±17.1] vs [81.3 ± 19.9]%), mid-expiratory flow (MEF) with 75% of forced vital ca-pacity (FVC) expired ([96.1 ±16.1] vs [88.8 ±23.1]%), MEF with 50%of FVC expired ([88.4 ±17.8] vs [84.8 ±20.8]%), and MEF with 25%of FVC expired ([92.7 ±25.9] vs [82.9 ± 28.7]%) (P<0.05), but had no statistically significant differences in the predicted percentages of FVC, forced expiratory volume in 1 second (FEV1), and the ratio of FEV1 to FVC (P>0.05).The positive rate of AHR was 6.1% (16/246) in the NAR group.All the indices of small airway function were significantly lower in the AHR than in the nAHR group (P <0.01). Conclusion NAR patients are apt to undergo obvious changes in small airway function, some with AHR, which is associated with lower airway function changes.